Single molecule traffic in mesoporous materials and new photostable water soluble terrylenediimide derivatives [Elektronische Ressource] / Christophe Jung

Christophe Jung

Sujets

• Biologie

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2007
• Nombre de lectures: 14
• Langue: English
• Poids de l'ouvrage: 33 Mo

Extrait

Dissertation zur Erlangung des Doktorgrades
de Fakult¨at fur¨ Chemie und Parmazie
der Ludwig-Maximilians-Universit¨at Munc¨ hen
Single Molecule Traffic in Mesoporous
Materials
AND
New Photostable Water-Soluble
Terrylenediimide Derivatives
Christophe Jung
from
Haguenau
2007Erkl¨arung
Diese Dissertation wurde im Sinne von § 13 Abs. 3 bzw. 4 der Promotionsordnung
vom 29. Januar 1998 von Prof. Dr. C. Br¨auchle betreut.
Ehrenw¨ortliche Versicherung
Diese Dissertation wurde selbst¨andig, ohne unerlaubte Hilfe erarbeitet.
Munc¨ hen, den 14. September 2007
Dissertation eingereicht am 14.09.2007
1. Gutachter: Prof. Dr. C. Br¨auchle
2. Gutachter: Prof. Dr. T. Bein
Mundlic¨ he Prufung¨ am 16.10.2007Contents
1 Introduction 5
2 Theoretical background and methods 9
2.1 Fluorescence microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2.1.1 Fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2.1.2 Single molecule spectroscopy (SMS) . . . . . . . . . . . . . . . . . 12
2.1.3 Apparatus and methods . . . . . . . . . . . . . . . . . . . . . . . 17
2.2 Diffusion theory and trajectory analysis . . . . . . . . . . . . . . . . . . . 33
2.2.1 Diffusion theory: microscopic approach . . . . . . . . . . . . . . . 33
2.2.2 Analysis of trajectories . . . . . . . . . . . . . . . . . . . . . . . . 35
3 Translational,orientationalandspectraldynamicsofsingledyemolecules
in highly oriented mesoporous materials 37
3.1 Introduction to mesoporous materials . . . . . . . . . . . . . . . . . . . . 37
3.2 Diffusion and orientation of single TDI molecules in different mesophases
of Brij-56-templated mesoporous films. . . . . . . . . . . . . . . . . . . . 42
3.2.1 Sample systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
3.2.2 Diffusionnal behavior of single TDI molecules in pure hexagonal
and lamellar phases . . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.2.3 Diffusion of single TDI molecules in the phase mixture - Phase
switching . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
12 CONTENTS
3.2.4 Mean squared displacement analysis. . . . . . . . . . . . . . . . . 51
3.2.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
3.3 Translational,orientationalandspectraldynamicsofsingleTDImolecules
in CTAB-templated mesoporous films . . . . . . . . . . . . . . . . . . . . 54
3.3.1 The M41S CTAB-templated mesoporous system . . . . . . . . . . 54
3.3.2 Simultaneous measurement of orientational and translational dy-
namics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
3.3.3 Simultaneous measurement of orientational and spectral dynamics 58
3.3.4 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
3.4 Preparation and single molecule investigation of highly structured do-
mains in CTAB-templated mesoporous films . . . . . . . . . . . . . . . . 74
3.4.1 Growth of the highly structured domains . . . . . . . . . . . . . . 74
3.4.2 Translational and orientational dynamics of single TDI molecules
diffusing in the highly structured domains . . . . . . . . . . . . . 85
3.4.3 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100
3.5 Experimental section . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
4 New photostable water-soluble terrylenediimide derivatives for appli-
cations in single molecule studies and membrane labelling 105
4.1 Introduction: novel fluorophores for biological labelling . . . . . . . . . . 105
4.2 WS-TDI as a powerful dye for applications in single molecule studies and
membrane labelling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
4.2.1 Absorption and emission spectra . . . . . . . . . . . . . . . . . . 109
4.2.2 Single molecule studies . . . . . . . . . . . . . . . . . . . . . . . 112
4.2.3 Protein labelling . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
4.2.4 WS-TDI as a membrane marker . . . . . . . . . . . . . . . . . . 117
4.3 Two other WS-TDI derivatives: WS-TDI dodecyl and WS-TDI pyridoxy 121CONTENTS 3
4.3.1 New derivatives for a larger panel of applications . . . . . . . . . 121
4.3.2 Photophysical parameters . . . . . . . . . . . . . . . . . . . . . . 121
4.3.3 Fluorescence spectra in presence in cells . . . . . . . . . . . . . . 126
4.3.4 Membrane labelling in living cells . . . . . . . . . . . . . . . . . . 127
4.4 Localization of the three WS-TDI derivatives in the early and late endo-
cytic pathway of living cells . . . . . . . . . . . . . . . . . . . . . . . . . 132
4.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
4.6 Experimental section . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138
5 Summary 141
5.1 Single molecule traffic in mesoporous materials . . . . . . . . . . . . . . . 141
5.2 Photostable water-soluble terrylenediimide derivatives for applications in
single molecule studies and membrane labelling . . . . . . . . . . . . . . 144
6 Appendix 147
6.1 Orientation of TDI molecules encapsulated in ordered mesoporous chan-
nels in anodic alumina membrane hosts . . . . . . . . . . . . . . . . . . . 147
6.2 List of abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
6.3 Published work and contributions to conferences and meetings . . . . . . 151
6.3.1 Publications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
6.3.2 Contributions to conferences and meetings . . . . . . . . . . . . . 1524 CONTENTSChapter 1
Introduction
Until20yearsagochemistrywasdealingwithmoleculesinthesenseofmoles. In1952E.
Schr¨odinger stated: ”...we never experiment with just one electron or atom or molecule.
In thought experiments we sometimes assume we do...” Hence, chemists were thinking
in terms of ensembles of molecules, nearly forgetting that each molecule is an individual
entity. Only slowly scientists started dreaming about resolving moleculeslly.
Thiswassupportedbythedevelopmentofmoreandmoresensitivedetectorsculminating
in high- performance detectors able to detect the fluorescence signal of single emitters.
In 1989 and 1990 the absorption and fluorescence excitation spectra of single molecules
1 2were measured for the first time by the groups of Mo¨rner and Orrit, respectively.
Those revealed that spectra of single molecules, which were until then considered as
a stable property of a given dye molecule, could exhibit unsuspected strong dynamics
such as intensity fluctuations or large spectral jumps. Since then a variety of single
molecule spectroscopy (SMS) methods have been developed to observe not only the
fluorescence spectra of individual emitters, but a whole range of properties like the
3–9orientation of their dipole moment, their photostability (i.e. their resistance against
10–14 15–21photo-destruction), or their translational diffusion. These techniques paved the
wayforanewtypeofinvestigation,wherenotonlythemeanvalueofaspecificmolecular
property can be obtained, but the distribution of values for this property with large
variations from molecule to molecule.
The advantage of SMS studies compared to standard ensemble techniques is crucial in
fields ranging from fundamental investigations up to industrial applications. A single
molecule incorporated as a guest into polymeric or porous solids, will act as a reporter
22–28revealing a wealth of details about the host structure and the dynamics involved.
A prominent example are mesoporous materials which constitute a rapidly growing field
of host systems. Their scientific and technological relevance is based on the presence
56
29,30of pores of controllable dimensions at the nanometer lengthscale. The presence of
voids enable these materials to discriminate between molecules and allow interactions
with only a subfraction of them. Research efforts in this field have been driven by
31–33 34emergingtechnologicalapplicationssuchasnanocatalysis, gasseparation, chemical
35 36,37 38,39sensing, photonics, or novel drug delivery systems. The current challenges
includethedesignofmesostructuresforoptimizedapplicationsaswellasthefundamental
understanding of host-guest interactions.
Standard characterization methods, such as NMR, IR, UV/Vis spectroscopy, electron
microscopy and X-ray diffractometry characterize the host structure and host-guest in-
40–42teractions at the ensemble level. However, the nanometer-scale behavior of the
molecules is both spatially and temporally heterogeneous, and a complete characteri-
zation of the host-guest system is not possible with these methods. Information about
such heterogeneities is indeed lost in the ensemble averaging process. Hence SMS tech-
niques are the methods of choice to directly observe details of the molecular behavior
because they work on a molecule-by-molecule basis. The position of a single molecule,
the orientation of its dipole moment as well as the position, intensity and width of the
fluorescence spectrum, or photophysical parameters such as brightness and photostabil-
ity provide valuable information about the host, which are difficult or even impossible
to obtain with standard methods.
The first part of this thesis is devoted to the detailed investigation via SMS of the
translational, orientational and spectral dynamics of single terrylenediimide (TD